Nixtamalization as a Method to Reduce Aflatoxin and Increase Available

Nutrients in Maize

International Maize and Wheat Improvement Center (CIMMYT)

El Batán, Mexico

Molly Carroll, 2018 World Food Prize Borlaug-Ruan Intern

Plymouth, Minnesota

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TABLE OF CONTENTS

Acknowledgements…………………………………………….………………..………….…2-4

Abstract…………………………………………………………………..…………...……….…5

Introduction ……………………………………………………………………………….…6-11

Role of Maize in Nutrition and Food Security…………………………..…………….8-10

Nixtamalization………………………………………………………………………10-11

Methods

I. Participants…………………………………………...……………………………11-12

II. Apparatus and Materials………………………………………………………..……12

III. Procedure……………………………..………………………………….………12-16

Nixtamalization and Tortilla Making……………..………………….14-16

Results………………………….…………………………………………………..….……..16-18

Discussion……………………………………………………………………….….……..…19-20

References………………………………………………………………………………....…21-23
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ACKNOWLEGEMENTS

This experience was truly inspiring, but it did not come without its challenges. The most

difficult being lack of experience in working in a lab and in the topics that were being

researched. I have never worked in a lab setting before, so it took some adjusting to understand

the reasoning and methods behind daily tasks. This required asking questions and asking for

help when I needed it. The purpose of my time at CIMMYT was to learn, and part of that was

learning how to manage tasks and take control of my success. Another challenge that came with

living in a different country was the language barrier. I knew very basic Spanish, but not to the

level necessary to understand technical scientific instruction or analysis. Thankfully my research

mentors spoke some English, but the majority of the researchers only spoke Spanish. This made

it difficult to understand some of the complex procedures and analyses we were conducting, but

it also gave me the opportunity to learn Spanish and how to communicate without solely relying

on words. Things did not always go as planned in the lab during my research, so I had to

problem solve and work with others to find or create a solution. The equipment that was the

primary data collection device was malfunctioning and did not produce consistent results;

Without this piece of equipment, the experiment could not be completed. Solving this issue took

creative thinking, patience, and understanding that it was acceptable to face challenges. This

realization came at a very opportune time because I faced many more interruptions. One of the

biggest logistical challenges I experienced was the window of delivery for the maize samples I
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would be analyzing. They were delivered very late in my internship, so I did not have the

opportunity to finish all the data collecting myself. Fortunately, I was not alone in the research

and the Maize Nutritional Quality team assisted me in every step including finishing collecting

data. My time in Mexico was a learning experience in every way, I gained new skills in

problem-solving, time management, organization, and gained a whole new worldview.

First and foremost, I would like to thank the late Dr. Norman Borlaug for sharing his

passion and commitment to solving the global hunger crisis by creating the World Food Prize

Foundation. This organization is a champion in showcasing what young people can do to create

change. I am forever grateful to have had the opportunity to be a Borlaug-Ruan International

Intern and carry on the legacy of Dr. Norman Borlaug. I would also like to extend my gratitude

to the Ruan family and Ambassador Kenneth M. Quinn for recognizing the potential of young

people as leaders in the fight against food insecurity.

Next, I would like to thank Ms. Crystal Harris for all her work in managing and

organizing the logistics of my internship at CIMMYT as well as the internships of the other 23

Borlaug-Ruan International Interns. I am so grateful for your tireless work in ensuring my safety

while I was abroad. I am so appreciative of you and the intern selection committee for

recognizing my passion and desire to create solutions to the global hunger crisis.

Tremendous thanks to Johanna Braun for making me feel welcome and ensuring my

internship ran smoothly and Natalia Palacios Rojas for serving as my mentor and guiding me

through the lab and my research. I would also like to thank Aide Liliana Molina Macedo and

the rest of the Maize Nutritional Quality team for assisting in the tasks of my research and
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extending their friendship. Thank you to CIMMYT staff in Mexico for graciously hosting me

and providing a home for the duration of my two months in Mexico.

Lastly, I must thank all of those who helped me in my journey to shape who I am today.

Thank you to my parents Maureen English Carroll and Ned Carroll for always supporting me

and ensuring I have the resources to explore and expand my passions. Thank you to my sisters

Kate and Bridget Carroll for being my forever role models and always pushing me to be the

best that I can. I am also grateful to my extended family and my friends for being a familiar

face to share my experiences with while abroad.

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ABSTRACT

Aflatoxin, a form of mycotoxin, is a secondary metabolite found in soil and organic

matter. They are produced by microfungi and have very detrimental effects on human and
animal health as well as food security. Contamination by aflatoxin is largely inevitable in crop
production, especially in drought conditions and intermediate temperatures. Aflatoxin are also
difficult to eliminate during food processing because of their resistance to heat, chemical and
physical treatments, and other widely used food processing methods. The most common strains
are Aspergillus flavus and Aspergillus parasiticus which produce four aflatoxin types: B1, B2,
G1, and G2, that are distinguished by the blue or green fluorescence under UV light. These
strains of aflatoxin are the most potent natural carcinogen. B1 produced by A. flavus will be
referred to as “aflatoxin” because it is the only strain present in the data. The versatility and ease
of maize, as well as the rich nutritional profile, makes it a go-to for many families in low-income
areas, especially where food insecurity and malnutrition are present. However, when
contaminated by aflatoxin, the nutrients in maize are quickly degraded. Nixtamalization as a
processing method for maize reduces the risk of disease by contamination and provides many
improved nutrition benefits to target the issue of food insecurity. This study aimed to determine
if nixtamalization was a practical and efficient method to reduce aflatoxin and increase nutrients
in maize native to Mexico. The results show that after nixtamalization, maize samples had
considerably less contamination than before. More research into nixtamalization as a process to
improve the nutrition of maize and reduce the presence of aflatoxin is important in the fight
against malnutrition and food insecurity in a growing world.
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KEY WORDS: Maize, Aflatoxin, Nixtamalization 


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INTRODUCTION

Mycotoxins are secondary metabolites produced by microfungi that have very

detrimental effects on human and animal health. Contamination by mycotoxins is a global

concern because of its unpredictable and unavoidable nature even with good processing and

storage practices. This paper will specifically discuss aflatoxins, a form of mycotoxin, and their

role in plant pathology. Aflatoxins are mainly found in soil and organic matter, which puts crops

at a high risk of contamination and therefore livestock and humans who consume the crops.

Health effects of aflatoxin are targeted in the liver, but they are also mutagenic, carcinogenic,

teratogenic, hepatotoxic, and have immunosuppressive effects (Bennett & Klich, 2003).

The four major aflatoxins are B1, B2, G1, and G2, they are distinguished based on the color

of their fluorescence under UV light, either blue or green. There are many strains of fungi that

produce aflatoxin, but the most common are Aspergillus flavus and Aspergillus parasiticus. A.

flavus is capable of producing only B1, B2, but A. parasiticus can produce all four types. These

specific strains of aflatoxin are considered to be the most potent natural carcinogen (Squire,

1981). B1 produced by A. flavus is the most common and broadly known aflatoxin, because of

this, the term “aflatoxin" will be used to refer to the B1 strand produced by A. flavus for the

remainder of this paper.

Aflatoxin contamination is highly variable in plants. Natural contamination occurs

commonly in cereal grains such as corn, soybeans, wheat, peanuts, and many other foods. This

contamination can occur before harvest or after harvest while in storage. Contamination before

harvest has been associated with drought conditions, and other factors that weaken plant

structure. The contamination after harvest is attributed to the poor storage environments of
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harvested crops that supply optimal conditions for mold and aflatoxin growth such as high

moisture content and humidity. The methods to prevent uncontrollable aflatoxin growth are

mostly preventative. This can include proper agricultural practices before, during, and after the

growing period, as well as drying of the crops after harvest to ensure the moisture content is low

enough to inhibit mold growth. Even with good practices, aflatoxin contamination is largely

inevitable and not easily eliminated during food processing. Aflatoxin are very resistant to heat,

chemical and physical treatments, and other widely used food processing methods (Alshannaq &

Yu, 2017).

Specifically discussing the aflatoxin plight in maize crops, areas with ambient

temperatures and dry conditions are most affected, such as the southern United States, Mexico,

India, and Africa. Aflatoxin can be easily identified with just the human eye by a yellow-green

or grey mold on the kernels, husks, and/or stems of maize crops. Along with being easily

identifiable, aflatoxin are easily spread. Aflatoxin are a saprophytic organism (obtaining energy

from dead and decaying organic matter) that spread by asexual spores the fungi produces. These

spores can be carried by the wind or insects and enter through any point on the crop that is

structurally compromised. Insects, including the European corn borer, sap beetle, and corn

earworm, can also provide direct entry of the spores into the plants. (Richard, 2007).

Contamination of grains used in animal feed can be carried through to animal products humans

eat such as eggs, milk, and meat. This presents a very difficult situation for developing nations

related to human health and food insecurity, but it also affects developed nations. There have

been proposed solutions for controlling the toxin manifestation in food and crops, however, like

the issue of food insecurity, no single solution exists. As stated by Brown et al. (1998), recent
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advances in research of plant breeding and gene editing/engineering have created the opportunity

to develop more fungi-resistant crops. This coupled with targeting genes engaged in regulatory

processes of mycotoxin development and the use of biocontrol agents have given an advantage to

farmers. Other postharvest prevention methods include detection and isolation of aflatoxin

contaminated food by government regulation and screening programs (Bennett & Klich, 2003).

However, because aflatoxin is a natural contaminant, none of these solutions have solved the

problem.

Role of Maize in Nutrition and Food Security

Maize, also referred to as “corn”, from a nutritional view, is an excellent source of

proteins, starch, lipids, and other nutrients. This crop is the major food source for one third of

the world’s population. Many undeveloped and developed nations such as Mexico, Africa,

Southeast Asia, Latin America, and parts of the United States, rely on maize as a primary food

(Nuss & Tanumihardjo, 2010). The versatility and ease of maize and its derived products makes

it a go-to for many families in low-income areas, especially where food insecurity is present. The

anatomy of a corn kernel has four primary structures, the pericarp, germ, endosperm, and tip cap.

The endosperm makes up 83% of the kernel and is where a majority of the starch is found with a

small amount of protein and vitamins and minerals. The germ is a large storage of fiber, protein,

iron, and zinc; and the pericarp and tip cap are the outer protective shields of the kernel. The

pericarp, germ, and tip cap make up 5%, 11%, and 1% of the corn kernel, respectively. (Vanara,

et al., 2018). This translates to about 72% starch, 10% protein, and 4% lipid (Inglett,1970).

Maize supplies an energy density of about 3.65 kcal/1 gram which is similar to that of other life
 Carroll | Page !10

sustaining crops; wheat yields 3.40 kcal/1 gram and rice yields 3.60 kcal/1 gram (USDA).

However, the nutritional quality of these staple crops can be significantly reduced with the

presence of mycotoxins, like aflatoxin.

Maize alone supplies an abundance of macro- and micronutrients, but the qualities of

some of these nutrients are disproportionate and deficient for human metabolic needs. Maize

alone cannot be relied on as a single source of food because it will lead to malnourishment. This

is becoming a large issue in underdeveloped, poor, and rural areas where maize is the only source

of consistent food that may be available. Some of these nutrients that are lacking in maize are

essential amino acids lysine and tryptophan, vitamins C, B and A, iodine, and iron. Research in

exogenous and endogenous maize fortification is being conducted to help improve macro- and

micronutrient quality and quality to combat malnourishment. This encompasses biofortification

of maize as well as making nutrients more bioavailable for absorption once consumed (Nuss &

Tanumihardjo, 2010). Different geographic regions require different fortification. For example,

provitamin A is most necessary in Zambia, Zimbabwe, Malawi, Colombia, Pakistan, Ethiopia,

Nigeria, Democratic Republic of the Congo, and Mali. Increased protein quality, with respect to

the deficient amino acids, is necessary everywhere in the world, but especially in South Africa;

and high kernel-zinc maize is urgent in Guatemala, Nicaragua, and Colombia (Rojas, et al.,

2016). Any degradation of the available nutrients by aflatoxin is detrimental to the communities

that rely on maize as a majority of their daily calories.

Globally, government aflatoxin regulation varies between countries. The regulated

maximum for aflatoxin by the United States Food and Drug Administration is 20 parts per billion
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(ppb) for humans and animals. In the European Union, the maximum is much lower, around

0.1-15ppb depending on the food being produced. In Mexico, the regulated value for aflatoxin is

30 ppb, but the actual value in various food products tests much higher values (USDA, 2009).

Maize can be consumed and prepared in a multitude of ways that vary around the world:

straight off the cob, boiled, ground, roasted, as animal feed, fermented, used in cakes, breads, and

alcoholic beverages, and nixtamalized. Further processing can also yield sweeteners, oils,

thickeners, and other by-products (Inglett,1970). As Nuss and Tanumihardjo (2010) state, “Maize

is a dietary staple for more than 200 million people. This number can be expected to grow as the

world's population approaches 8 billion in 2025, indicating maize's status as a paramount crop in

the context of global nutrition.” The importance of maize will only increase as the number of

people needing nutritious foods increases, so research into the aflatoxin and nutrient fortification

are necessary to feed the growing world.

Climate change is changing how the agriculture industry functions worldwide. Shifts in

temperature, weather patterns, and other factors are creating and destroying growing areas

resulting in loss of efficiency, crop yield, and income (Stepman, 2018). This change also

increases the likelihood of pathogenic activity that could dramatically affect the economic and

social systems in the agriculture industry (Magan, et al.) (Battilani, 2016).

Nixtamalization

Nixtamalization is a processing method for maize to increase nutritional value, decrease

aflatoxin levels, and prepare the maize for food products. In a traditional nixtamalization

process, maize is cooked and steeped in a solution of lime (calcium hydroxide) and wet-milled to
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produce masa. The lime concentration usually contains 0.5-3% of calcium hydroxide mixed with

water to create an over saturated solution. Part of the nixtamalization process requires washing

and cooking of the maize. The maize is added to the correct solubility of lime solution

(depending on the amount of maize), cooked in an open vat, and then left to steep overnight. The

maize is then washed and the pericarp, which is where most of the aflatoxin reside, is manually

removed from the kernels. The name ‘Nejayote’ is given to the liquid after cooking that contains

pericarp fragments, remaining lime, and water. During this process, the maize kernels swell to

about 1.5 times their original size; 28-30% of water absorption is done during cooking, and 5-8%

during steeping. The amount of water absorbed is dependent on the kernel size, pericarp

thickness, lime concentration, and the type of endosperm (McDonough, et al., 2001).

Nixtamalization provides many improved nutrition benefits that can reduce the malnourishment

issue when maize is the primary food in a diet. These nutrition and health benefits include:

decreasing the risk of the disease pellagra by increasing niacin bioavailability, increasing calcium

levels from absorption of the kernels during steeping with the calcium hydroxide solution,

increasing resistant starch which supplies dietary fiber; and substantially decreases mycotoxin

levels. Along with health benefits, nixtamalization also lengthens the shelf life of food products

and creates businesses and market opportunities for income in a community (Rojas, et al., 2016).

This process yields a dough called masa which is used in over 300 food products in Mexico, the

most recognized being tortillas (Moctezuma-Zárate, 2015). More research into nixtamalization

as a process to improve the nutrition of maize and reduce the presence of aflatoxin is important

in the fight against malnutrition and food insecurity in a growing world. This study aimed to

determine if nixtamalization is a practical food safety processing method for maize in the
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household to industry setting; as well as how efficient nixtamalization is at reducing aflatoxin.

Given the background into the anatomy of a maize kernel and aflatoxin, nixtamalization has

great potential to decrease aflatoxin in maize varieties native to Mexico while increasing

available nutrients.

METHODS

I. Participants

Enrolled in this experiment was maize from the southern and central regions in Mexico.

Each sample of grain was treated with a chemical control agent, but that was not a relevant

variable to this study. The color of the grain also varied from white to yellow, however this was

also not a relevant variable. The color of maize is mostly based on preference for consumers.

The maize samples were only analyzed for aflatoxin levels.

II. Apparatus and Materials

To collect the data in this experiment, the devices used were designed specifically for use

in quantifying aflatoxin. A transportable box with a ultraviolet light was used to identify if there

was a presence of aflatoxin in the grain sample. This can only be used to identify which samples

were contaminated by aflatoxin, but a chemical analysis of the grain is needed to quantify the

amount present. An analytical balance scale was also used to weight samples of grain for

analysis. The precision and accuracy of this device was necessary for the detail and accuracy of

this study. A cold storage room was used to store the samples of grain at -15°C to inhibit the

growth of aflatoxin. This was necessary to ensure the amount of aflatoxin present on the grain

samples was from the growing or harvesting period, not from storage and analysis in the lab.
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Various millers were also used to grind the maize samples to different sizes. In order to quantify

the aflatoxin with the equipment available the grain needed to be ground to at least 1.5

millimeters in diameter. The final apparatus was AccuScan. This equipment consisted of small

strips, similar to pH strips, that absorbed aflatoxin in a sample and measured it in a handheld

device. This equipment was the primary apparatus used to collect data for this study.

III. Procedure

To begin this experiment, each sample of corn was assigned a sample number; the

numbers were assigned randomly based on the position of the sample bag in the pallet. The

numbers ranged from 4854 to 4949 as a continuation of the numbers from past experiments in

the lab. The purpose of the number was to easily track the samples during experiments to

connect with background data in analysis. Once numbered, each sample was tested under

ultraviolet light to check for the presence of aflatoxin. The sample was mixed to homogenize it

and poured onto a tray so the bottom was fully covered. The tray was placed in a portable UV

box and the number of grains that glowed due to aflatoxin contamination were counted and

recorded; three trials were performed for each sample. To prepare for the preliminary chemical

analysis of the grain, the samples were shaken to homogenize the maize and 500 grams of grain

from each sample were weighed using an analytical balance scale. These samples were milled to

produce flour at 0.5 millimeter and 1.5 millimeter thickness for each sample. The different

thicknesses was an independent variable to determine if aflatoxin levels tested differently. 20

grams of the milled flour from each sample was weighed using an analytical balance scale and

added to sample bottle labeled with the appropriate sample number. To this, one packet of MAX
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1-G50 aqueous extraction was added from the AccuScan kit as a precursor for analysis. To

create a solution, 50 millimeters of distilled water was added to each sample bottle with the

maize flour and placed in a vertical sample rocker to vigorously shake the samples for three

minutes. The consistent amount of flour and water measured for each sample analysis was a

controlled variable. Once the flour was completely mixed with the water, the samples were left

to rest for five minutes to separate and form distinct layers based on particle size; Only the top

layer (smallest particle size) is needed for this analysis. The solution was carefully poured, so

only the top layer is transferred, into the top of a syringe with a cotton ball at the base for

filtration, and filtered into a labeled test-tube. 100 microliters of sample diluent (provided by the

AccuScan kit) was pipetted into red sample cups. Before beginning analysis, the handheld

AccuScan Gold reader (provided by AccuScan kit) needed to be programed to the “Mycotoxin

Q+ MAX” category and specifically the “Afla Q+ MAX” test to analyze aflatoxin level. Once

the reader was programed, 100 microliters of the sample extract was placed in a red sample cup

(also labeled with the correct sample number) with the sample diluent and mixed five times

using the pipette. After mixing, one “Reveal Q+ MAX for Aflatoxin” test strip was placed in the

sample cup, so the end of the strip came in contact with the solution and could begin to wick, for

six minutes. After the allotted development time, the strip was removed and placed into the

AccuScan Gold reader to quantify the aflatoxins present. This procedure relied on following the

procedure correctly and without error to ensure the correct results were obtained. This analysis

was repeated for each sample (4854 to 4949). The dependent variable was the aflatoxin level,

measured in parts per billion (ppb) for each sample. If the aflatoxin result was higher than

50ppb, the AccuScan could not provide an accurate reading, so the sample extract needed to be
 Carroll | Page !16

diluted. A dilution by six times was the first dilution and was made using 500 microliters of

distilled water and 100 microliters of the sample extract. The chemical analysis was preformed

again and if the result was higher than 50ppb, an extraction by seven times was made and tested;

this continued until the result was under 50ppb to ensure a correct quantification of aflatoxin in

the sample. To counteract the dilution in the results, the aflatoxin number was multiplied by the

amount it was diluted by; for example, a result of a dilution by six times was multiplied by six.

Nixtamalization and Tortilla Making:

To begin the nixtamalization process, un-milled maize grain was added to a cooking pot.

The weight of each sample varied based on how much total sample was available, but the

average weight was around 600 grams. The calculations of distilled water and lime (calcium

hydroxide) were dependent on the initial weight of the grain sample. The amount of distilled

water added to a sample (in milliliters) was double the weight of the sample. The amount of lime

added was dependent on what solubility or percent concentration was necessary, for this trial 1%

was needed, so the weight of 1% of the grain sample was measured in lime. The solution formed

was alkaline with a pH of around 11. The pots with samples were then cooked for 35 minutes at

a constant temperature of 80°C and then left to steep (off the heat) for around 16 hours. After

steeping, the grain was strained and the Nejayote (liquid from straining) was collected and

labeled with the grain sample number for chemical analysis of aflatoxin. The grain, now called

nixtamal, was rinsed twice and during the first rinse, the pericarp on the grain was manually

removed by rubbing the kernels and creating a disturbance. The “clean” grain was then put into

a wet-miller and ground with water to create masa. The control variables were the percentage of
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lime, proportions of reactants, time for cooking and steeping, and temperature for cooking. This

process was repeated for each sample.

The masa produced from nixtamalization was then used to make tortillas. The masa is

kneaded with water to the correct consistency needed for traditional tortilla making. 20 grams of

the masa dough was weighed using an analytical balance scale and flattened using a traditional

tortilla press to achieve the correct shape and thickness needed for cooking. The fattened dough

was then placed onto a hot tortilla pan (on a stove) and cooked for approximately 17 seconds on

each side. This process was repeated 20 times for each sample. Once the tortillas were made,

they were shredded into smaller pieces (to create more surface area) and placed into a flash

freezer to preserve the tortilla. Once completely frozen, the tortilla pieces were placed into a

freeze drier to rid of any moisture. After frozen, the tortilla pieces were milled to both 0.5mm

and 1.5mm thickness. The chemical analysis described above was repeated for the nixtamalized

samples after adjusting the moisture level. Moisture was adjusted by adding water to the milled

tortilla and mixing; the moisture after this process was between 10-12%.

RESULTS

This study aimed to determine if nixtamalization could be used as a food safety

processing method to improve the nutrition of maize and reduce the presence of aflatoxin. This

research is important in the fight against malnutrition and food insecurity in a growing world.

Given the background into the anatomy of a maize kernel and aflatoxin, nixtamalization has the

potential to serve as a life changing practice in food sustainability and human health. The

primary measurement was in aflatoxin levels of maize samples from Mexico. Quantification of
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aflatoxin was determined at each stage of nixtamalization; the grain, nixtamal, nejayote, masa,

and tortillas; and compared to determine how efficient nixtamalization is as a methods to

improve food safety. Each sample was nixtamalized at 1% and 1.5% lime (calcium hydroxide)

concentration to test whether this variable had an effect on aflatoxin levels. The output of masa

from nixtamalization was further cooked as a tortilla and the tortilla was also analyzed for

aflatoxin levels. The changes in heat and pH during nixtamalization and cooking were of interest

to observe how it effects aflatoxin.

Figure 1. compares the

percent of average grains
contaminated found in
the UV light test to
aflatoxin quantification
values found by
performing chemical
analysis.

The ultraviolet light analysis results of the grain samples show that there is a relationship

between the amount of fluorescence and level of aflatoxin. While the test did not provide results

on the qualification of aflatoxin, it was able to accurately predict the aflatoxin range based on

percentage of grain contaminated. The samples with a similar percent of average grains that

Table 1. Displays the values from aflatoxin qualification of the outputs from nixtamalization:
including raw grain, nixtamal, nejayote, masa, and tortillas.
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showed fluorescence had similar values when the chemical analysis for aflatoxin was performed

The results of the nixtamalization section of this study show that overall the process of

nixtamalization dramatically reduces aflatoxin levels in the maize samples that were tested. The

values show that some of the aflatoxin was distributed to the najayote in the form of pericarp

from the maize. It also shows that the cooking process from raw grain to nixtamal and from

masa to tortilla had very little effect on aflatoxin level; this was expected because aflatoxin are

very resistant to heat. When discussing the results of the two different lime concentration, both

concentration effectively reduced aflatoxin in the maize samples, but 1.5% had a higher

Figures 2 & 3. Display the aflatoxin values from the outputs of

nixtamalization in graphical form for ease of analysis.
 Carroll | Page !20

reduction in aflatoxin than 1%. The higher lime concentration caused more aflatoxin to be

transferred to the najayote which resulted in lower aflatoxin in the nixtamal.

Figure 2 & 3. Displays the values from aflatoxin qualification of the outputs from nixtamalization:
including raw grain, nixtamal, nejayote, masa, and tortillas in a graphical form for ease of analysis.

DISCUSSION
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From the results of this study, it can be concluded that nixtamalization is an effective

method to reduce aflatoxin in maize. Nixtamalization also increases nutrient availability to make

maize a more nutritiously well-rounded staple crop. This conclusion is important as a possible

solution to malnutrition and food instability in low-income areas. Maize is heavily relied on for

a majority of daily nutrients and calories, so determining a way to make this crop have a larger

impact would have beneficial effects on countless populations of people. The large reduction in

aflatoxin by the 1.5% lime solution shows that a higher concentration of lime, and therefore a

lower pH, is needed to combat the high stability of aflatoxin. Heat did not seem to have a

significant effect on aflatoxin because of its high resistance, but further research into a range of

temperatures is needed to make a confident conclusion.

Results of similar studies show results of the same nature. There have been very few

studies done to compare aflatoxin levels in maize before and after nixtamalization, but the ones

that have been published have come to comparable conclusions. The differences between this

study and others lie in the samples tested. There is a large focus on maize in Africa, which is

slightly different than the maize varieties native to Mexico that were analyzed in this study.

Aflatoxin is also highly variable in analyses, this can create more differences in results

depending on how procedures for analysis were performed.

There were many limitations in this study. The primary limitation was time. This

research had a very small window to complete data collection which made it difficult because the

procedure required a lot of time for each sample. Another limitation was in the aflatoxin itself,

aflatoxin are highly variable when performing analysis, so one test of a sample could produce
 Carroll | Page !22

different results if performed twice. This required each analysis to be diluted or performed again

and greatly reduced efficiency. The equipment used in this study was also a limiting factor. The

aflatoxin analysis equipment was not functioning properly which required data collection to be

halted until the equipment was fixed. Similarly, a piece on the wet-miller equipment used in

grinding the nixtamal into masa was broken and required data collection to stop until it was

fixed.

This study raises new questions for future work and other solutions to food insecurity.

Based on the results of aflatoxin after nixtamalization in different lime concentrations, it would

be beneficial to continue testing at different concentrations to determine which is optimal for

aflatoxin removal. Nixtamalization, or a process similar, could also be used on different grains

contaminated by aflatoxin to discover if this process is effective for other crops. Alternative

methods for removal of the pericarp on maize kernels could also be studied because a majority of

aflatoxin reside in and around the pericarp. The final question this study raises is how to make

nixtamalization a widely used food safety process that is accessible in homes. The results show

that nixtamalization is effective at removing aflatoxin from maize, but in order for these results

to have an effect on populations, the methods need to be practical for everyday use. Utilizing

nixtamalization, malnutrition and food insecurity in populations would start to diminish and the

quality of life would drastically increase.

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